CN115624517A

CN115624517A - Probiotic metaplasia product and application thereof in promotion of skin health

Info

Publication number: CN115624517A
Application number: CN202211328785.6A
Authority: CN
Inventors: 其木格苏都; 郭霄; 刘晓军; 包维臣
Original assignee: Inner Mongolia Ketuo Biology Co ltd
Current assignee: Inner Mongolia Ketuo Biology Co ltd
Priority date: 2022-10-27
Filing date: 2022-10-27
Publication date: 2023-01-20

Abstract

The invention discloses a probiotic metaplasia product and application thereof in promoting skin health, wherein the probiotic metaplasia product comprises: inactivated thallus and metabolite thereof after fermentation of probiotics; the probiotics at least comprise lactobacillus paracasei Probiosci-101 and lactobacillus fermentum ProSci-602; the preservation number of the lactobacillus paracasei Probiosci-101 is CGMCC No.25085, and the preservation number of the lactobacillus fermenti ProSci-602 is CGMCC No.25451. The invention can obtain the metabolite which can effectively improve the skin state through the mutual matching of 2 strains, and has obvious effect.

Description

Probiotic postbiotic product and application thereof in promotion of skin health

Technical Field

The invention relates to the technical field of biology, in particular to a probiotic postbiotic product and application thereof in promoting skin health.

Background

The skin is the largest organ of the human body, and about one billion microorganisms live on the skin, that is, more than two million microorganisms live on one piece of skin about the size of one rouleau coin. About 74-80% of these microorganisms are bacteria, 5-10% are fungi, and 10-20% are phages (natural enemies of bacteria), and a delicate balance is maintained between different species. The balance is not only different from person to person, but also influenced by environment, climate, age and living habits all the time, changes ceaselessly and records the experience of the whole life faithfully, and people with the same distribution of two colonies cannot be found in the world like genes and fingerprints. As a balance, our microecology of the skin surface is constantly experiencing various challenges, and studies have shown that: the micro-ecology of the skin is susceptible to environmental pollution and aging. The environmental pollution is likely to introduce foreign microorganisms, so that the skin micro-ecology becomes more and more complex and loses balance; the decrease of sebum secretion caused by aging, especially around menopause, changes the decrease of the proportion of microorganisms that feed on the skin and changes the proportion of the equilibrium state. The most common consequences of skin dysbiosis are acne, skin irritation, and dry scaling.

The metazoan is a product obtained by directly culturing probiotics in vitro and metabolizing, can directly generate positive effect on skin, and is the most common microecological technical means in skin care products nowadays. Survey data has shown that the proportion of women who listen to and purchase skin care products containing "probiotic" ingredients is as high as 66%, indicating that the use of probiotics has become widely accepted by the prebiotics. However, although the metabolites of the probiotic post-biotic products disclosed in the prior art can achieve the purposes of maintaining the skin micro-ecological balance, regulating the oil and water balance and further improving the skin state, the improvement effects are not very obvious, and especially the effects on improving skin melanin and skin wrinkles are not ideal.

Disclosure of Invention

Therefore, the technical problem to be solved by the present invention is to overcome the defect that the skin improvement effect of the probiotic post-biotic product disclosed in the prior art is not very obvious, so as to provide a probiotic post-biotic product capable of more effectively improving the skin condition and an application thereof in promoting skin health.

A probiotic post-biotic product comprising: inactivated thallus and metabolite thereof after fermentation of probiotics; the probiotics at least comprise lactobacillus paracasei Probiosci-101 and lactobacillus fermentum ProSci-602; the preservation number of the lactobacillus paracasei Probiosci-101 is CGMCC No.25085, and the preservation number of the lactobacillus fermenti ProSci-602 is CGMCC No.25451.

The Lactobacillus paracasei Probiolci-101 contained in the product is separated from Xinjiang Pamilar yoghurt, is preserved in the China general microbiological culture Collection center (CGMCC No. 25085), is classified and named as Lactobacillus paracasei Paracelasei, has the preservation time of 2022 years and 6 months and 13 days, has the preservation address of No. 3 Haematole No. 1 Xilu of the Chaxingyuan Shangyi of Beijing, and is detected as a viable strain by the preservation center at 2022 years and 6 months and 13 days. The strain has good capability of resisting gastric acid and bile salt environments; can improve lipid metabolism, reduce blood lipid, and balance immunity.

The Lactobacillus plantarum ProSci-602 contained in the product is separated from breast milk of healthy women, is preserved in the China general microbiological culture Collection center (CGMCC) No.25451, is classified and named as Lactobacillus fermentum, is preserved for 8 months and 1 day at 2022 years, is preserved for 3 days at Xilu 1 north Chen of the sunward area in Beijing, and is detected as a viable strain at 8 months and 1 day at 2022 by a preservation center. The experimental study proves that: the lactobacillus fermentum ProSci-602 has good capability of tolerating gastrointestinal fluids and bile salts, and can prevent colonization and infection of latent pathogenic bacteria in oral cavity such as dental caries bacteria.

The metabolite contains more than 50mg/g of organic acid, more than 7mg/g of short-chain fatty acid, more than 2mg/g of amino acid, more than 90mg/g of micromolecule polypeptide and more than 7mg/g of active substance with bacteriostatic efficacy. Preferably, the polypeptide comprises more than 100mg/g of small molecule polypeptide.

The organic acid comprises one or more of citric acid, succinic acid, tartaric acid, malic acid, 4-hydroxy phenyl lactic acid, phenylalanine, benzoic acid and oxalic acid;

the short chain fatty acids comprise one or more of lactic acid, propionic acid, butyric acid, caproic acid and acetic acid;

the amino acid comprises one or more of valine, tyrosine, proline, phenylalanine, leucine, isoleucine, alanine, aspartic acid, glutamic acid, lysine and histidine;

the active substance comprises: one or more of arachidic acid, methoxychalcone, tacrolimus, rokitamycin, isostearic acid, marnolide, caffeic acid, pinoresinol, qingdainone, ceramide, astaxanthin, gibberellin, schisandrin, ergosterol, tylosin, vitamin B2, rhein, limoflavin, ecdysterone, erythrulose, theophylline, benzyl cinnamate, salvianolic acid, hydrocortisone, squalamine, homosalate, piperlonguminine, and retinoic acid.

The content of the citric acid is more than 50 mg/g;

the content of the lactic acid is more than 5mg/g, and the content of the caproic acid is more than 1.5 mg/g;

the content of valine is more than 0.6mg/g, the content of tyrosine is more than 0.4mg/g, the content of proline is more than 0.3mg/g, and the content of phenylalanine is more than 0.2 mg/g;

the content of dipeptide in the small molecular polypeptide is more than 30mg/g, the content of tripeptide is more than 50mg/g, the content of tetrapeptide is more than 10mg/g, and the content of pentapeptide is more than 0.5 mg/g;

the content of the arachidic acid is more than 2mg/g, the content of the methoxychalcone is more than 0.9mg/g, the content of the tacrolimus is more than 0.8mg/g, the content of the rokitamycin is more than 0.8mg/g, the content of the isostearic acid is more than 0.8mg/g, the content of the marnolide is more than 0.6mg/g, the content of the caffeic acid is more than 0.2mg/g, and the content of the pinoresinol is more than 0.1 mg/g.

The ratio of the colony numbers of the lactobacillus paracasei Probiosci-101 and the lactobacillus fermentum ProSci-602 is 2-3;

preferably, the ratio of the number of colonies of lactobacillus paracasei Probiosci-101 and lactobacillus fermentum procci-602 is 1.

The culture medium used for fermentation comprises the following components in percentage by mass: 1.5 to 7 percent of full-fat soybean powder, 11 to 18 percent of skim milk powder, 0.05 to 0.5 percent of sodium citrate and the balance of water.

The product is a cosmetic, preferably a functional cosmetic.

A method of preparing a probiotic post-biotic product comprising:

obtaining a probiotic strain and a culture medium in the probiotic post-biotic product;

inoculating probiotic strains into the sterilized culture medium, fermenting at constant temperature until the pH is 4.5-4.6, inactivating, and drying to obtain probiotic post-production product.

The temperature of the constant-temperature fermentation is 33-37 ℃.

The culture medium further comprises a homogenization step after the completion of the preparation and before the sterilization;

preferably, the homogenizing temperature is 55-60 ℃, and the pressure is 18-20.0 Mpa;

the temperature of the culture medium during sterilization is 95 ℃, and the sterilization time is 60min;

the method also comprises a homogenizing step after fermentation and before inactivation, wherein the homogenizing temperature is 55-60 ℃, the primary pressure is 18-20.0 Mpa, and the secondary pressure is 5.0Mpa;

the drying is spray drying.

The probiotic post-biotic product or the application of the probiotic post-biotic product prepared by the preparation method in promoting skin health.

The application for promoting skin health includes but is not limited to the application in any one of the following:

1) Auxiliary prevention and/or improvement of facial skin condition;

2) The application of the product in preparing products for assisting in preventing and/or improving the facial skin state;

the skin condition includes, but is not limited to, increasing facial elasticity, reducing facial wrinkles, increasing facial moisture content, reducing facial oiliness, reducing facial melanin.

The technical scheme of the invention has the following advantages:

the invention provides a probiotic metaproduct, which optimizes probiotic strains for fermentation, selects the probiotic strains with different sources, different types and different probiotic characteristics to cooperate with each other, and the probiotic strains in the invention at least comprise Lactobacillus paracasei Probios ci-101 and Lactobacillus fermentum ProSci-602; through the mutual cooperation of the at least 2 strains, a culture substrate can be more effectively converted into a metabolite which is more beneficial to improving the skin state, the application effect of the probiotic post-production product in improving the skin health is improved, particularly, the aim of conventionally adjusting microbial flora on the skin can be fulfilled, the water-oil balance is achieved, the effects of increasing the facial elasticity, reducing facial wrinkles, reducing facial melanin and the like can be effectively achieved, and the comprehensive effect is very excellent.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a graph showing the moisture content level of the skin of volunteers in Experimental example 2 after 28 days of life span using probiotics; in a figure: the used skin care product for promoting health is used for 0 day and 28 days; and (b) in the figure: using the after-growth skin care product with or without the skin care product for 28 days;

FIG. 2 is a graph showing the skin elasticity of volunteers in Experimental example 2 after 28 days of life with probiotics; in a figure: the used skin care product for promoting health is used for 0 day and 28 days; and (b) figure: using postbiotic skin care products with and without postbiotic skin care products for 28 days;

FIG. 3 is the skin melanin levels of volunteers in Experimental example 2 after 28 days of life span with probiotics; a, drawing a: the used skin care product for promoting health is used for 0 day and 28 days; and (b) figure: using the after-growth skin care product with or without the skin care product for 28 days;

FIG. 4 is the level of skin oiliness 28 days after probiotic application by volunteers in Experimental example 2; in a figure: the used skin care product for promoting health is used for 0 day and 28 days; and (b) in the figure: using the after-growth skin care product with or without the skin care product for 28 days;

FIG. 5 is a graph showing the wrinkle level of volunteers in Experimental example 2 after 28 days of prebiotics use of probiotics; in a figure: after the skin care product is used, the skin care product is born for 0 day; and (b) figure: after being used, the life-prolonging skin care product is used for 28 days; and (c) figure: the used skin care product for promoting health is used for 0 day and 28 days; FIG. d: postbiotic skin care products with and without postbiotic skin care products were used for 28 days.

Detailed Description

Example 1

A probiotic post-biotic product is prepared by the following steps:

1) Weighing the culture medium: mixing 5% of whole bean powder, 15% of skim milk powder, 0.25% of sodium citrate and 79.75% of water according to the proportion requirement;

2) Material melting: melting the materials at 58 ℃ to obtain feed liquid;

3) Homogenizing: at 58 deg.C and 19.0MPa;

4) And (3) sterilization: sterilizing the homogenized material liquid of 3) at 95 deg.C for 60min;

5) And (3) cooling: cooling the sterilized feed liquid to 35 ℃;

6) Adding strains and lactase: adding strains with viable bacteria number ratio of 1 to the feed liquid, wherein the total inoculation amount of the strains in the feed liquid is 4 multiplied by 10 for lactobacillus paracasei Probiosci-101 and lactobacillus fermenti ProSci-602 ⁶ CFU/mL. Adding 0.5g/1000g of lactase according to production requirements, and providing a carbon source for probiotics through enzymolysis;

7) Constant-temperature fermentation: performing constant temperature fermentation at 35 deg.C until pH is 4.5-4.6, wherein the fermentation step is about 18-19 hr (18 hr in this example), and the final pH value is 4.6;

8) Inactivation: mixing maltodextrin according to production requirements (providing ideal liquid dispersion for spray drying), preheating to 58 deg.C, homogenizing under first-stage pressure of 19.0Mpa and second-stage pressure of 5.0Mpa, sterilizing and inactivating at 75 deg.C for 15min;

9) Spray drying: and (3) spray-drying the inactivated bacterial suspension (the air inlet temperature is 160 ℃, and the air outlet temperature is 45 ℃) to obtain the probiotic post-vitality powder.

Example 2

A probiotic preparation, this example differing from example 1 in the composition of the culture medium, which in this example is: 1.5 percent of full-fat soybean powder, 18 percent of skim milk powder, 0.05 percent of sodium citrate and the balance of water.

The material melting temperature in the step 2) is 55 ℃;

the homogenizing temperature in the step 3) is 55 ℃, and the pressure is 20.0Mpa;

the fermentation time in the step 7) is 19h, and the pH value at the end point of the fermentation is 4.5;

the homogenization temperature in the step 8) is 60 ℃, and the primary pressure is 18.0Mpa.

Example 3

A probiotic preparation, this example differing from example 1 in the composition of the culture medium, which in this example is: 7% of full-fat soybean powder, 11% of skim milk powder, 0.5% of sodium citrate and the balance of water.

The material melting temperature in the step 2) is 60 ℃;

the homogenizing temperature in the step 3) is 60 ℃, and the pressure is 18Mpa;

the homogenization temperature in step 8) was 55 ℃ and the primary pressure was 20.0MPa.

Example 4

A probiotic post-biotic preparation, this example differing from example 1 in that the ratio of the number of colonies of lactobacillus paracasei Probiosci-101 and lactobacillus fermentum proci-602 was 2.

Example 5

A probiotic post-biotic preparation, this example differing from example 1 in that the ratio of the number of colonies of lactobacillus paracasei Probiosci-101 and lactobacillus fermentum proci-602 was 3.

Test example 1

The probiotic post-biogenesis powder prepared in examples 1-5 is respectively subjected to LC-MS measurement to determine metabolites contained in the probiotic post-biogenesis. Since the detection means of each component is conventional, the specific detection process is not described herein again.

The determination results of the probiotic post-production powder obtained in examples 1 to 5 are shown in tables 1 to 3 below. Table 1 shows the results of the determination of organic acids and short chain fatty acids in the probiotic powders of examples 1-5; table 2 shows the results of the determination of amino acids and small molecule polypeptides in the probiotic post-prebiotic powders of examples 1-5, and table 3 shows the results of the determination of active substances in the probiotic post-prebiotic powders of examples 1-5.

TABLE 1

TABLE 2

TABLE 3

From the above table, it is clear that the contents of the components detected in examples 2 to 5 are not significantly different from those in example 1.

Test example 2

The effect of the probiotic post-biotic powder prepared in example 1 on improving skin health was verified, specifically:

1 test grouping

Randomly selected 110 (60 women, 50 men) subjects without skin disease and skin infection, and aged between 25-28 years. All volunteers controlled not to use other cosmetic products during the first 1 week (week 0 to week 1). The groups were randomly divided into metazoan groups and control groups. The provided skin-beautifying products (prebiotic group: sleeping mask cream group added with 1% probiotic prebiotic powder; control group: non-prebiotic mask cream group) were used in weeks 2 to 5, respectively. Within 8 hours prior to sampling, the subject was asked to refrain from rinsing with anything (including water), otherwise typical routine hygiene procedures were allowed. The two groups of products are all washing-free sleeping mask cream, and the using method of the product is as follows:

(1) When the noodles are cleaned at night, oil stains and cosmetics in pores are cleaned;

(2) Taking a proper amount of the product, and uniformly coating the product on the face in a light and thin manner to avoid the area around eyes;

(3) After the face is smeared, the face is stood for 20 minutes and is gently massaged until the face is absorbed;

(4) After absorption, the user can fall asleep directly without washing or after washing.

0. The skin phenotype (skin moisture content, skin elasticity, skin melanin, skin oiliness, skin wrinkles) of the face of the subject was examined on day 28 using dermaphanidermaphaeresis specialist Derma-Expert MC760, respectively.

2 results of the experiment

The gas chromatography detection result shows that: the anabiotic group is obviously superior to the control group in the aspects of skin moisture content, skin elasticity, skin melanin, skin oiliness and skin wrinkle improvement. The test results are shown in fig. 1-5, specifically:

the results of analysis of the skin moisture content measurement indicators are shown in fig. 1, and the skin moisture content levels of the metazoan group and the control group both tended to increase after treatment, but the metazoan group increased more significantly (p < 0.01).

As shown in fig. 2, the analysis results of the skin elasticity measurement index showed that the metazoan group and the control group both tended to increase in skin elasticity after treatment, but the metazoan group increased more significantly (p < 0.01).

The results of analysis of skin melanin detection indexes are shown in fig. 3, the metazoan group has a significant effect (p < 0.01) on reducing the skin melanin level, and the melanin level of the control group is increased after 28 days of treatment.

The results of analysis of the skin oiliness index are shown in fig. 4, and the metazoan group and the control group both have a tendency to decrease after treatment in terms of skin oiliness, but the metazoan group decreases more significantly (p < 0.01).

As shown in fig. 5, the analysis results of the skin wrinkle detection index showed that the metazoan group and the control group both tended to decrease in the skin wrinkles after the treatment, but the metazoan group decreased more significantly (p < 0.01).

3 conclusion

Through the detection and analysis of the facial skin level of the subject, the metazoan can achieve the effect of repairing the skin by reducing the skin melanin, oiliness and wrinkle level (p < 0.01), and increasing the skin elasticity and the skin moisture content (p < 0.01). Human body tests show that the product has obvious effect on repairing skin.

It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.

Claims

1. A probiotic post-biotic product, comprising: inactivated thallus and metabolite thereof after fermentation of probiotics; the probiotics at least comprise lactobacillus paracasei Probiosci-101 and lactobacillus fermentum ProSci-602; the preservation number of the lactobacillus paracasei Probiosci-101 is CGMCC No.25085, and the preservation number of the lactobacillus fermenti ProSci-602 is CGMCC No.25451.

2. The post-probiotic prebiotic product of claim 1 wherein the metabolite comprises 50mg/g or more of an organic acid, 6mg/g or more of a short chain fatty acid, 2mg/g or more of an amino acid, 90mg/g or more of a small molecule polypeptide, 7mg/g or more of an active substance.

3. The probiotic post-biotic product according to claim 2 wherein the organic acid comprises one or more of citric acid, succinic acid, tartaric acid, malic acid, 4-hydroxyphenyllactic acid, phenylalanine, benzoic acid and oxalic acid;

the active substance comprises: one or more of arachidic acid, methoxychalcone, tacrolimus, rokitamycin, isostearic acid, marnolide, caffeic acid, pinoresinol, qingdainone, ceramide, astaxanthin, gibberellin, schizandrin, ergosterol, tylosin, vitamin B2, rhein, limonin, ecdysterone, erythrulose, theophylline, benzyl cinnamate, salvianolic acid, hydrocortisone, squalamine, homosalate, piperlonguminine, and retinoic acid.

4. The probiotic post-biotic preparation according to claim 3,

the content of the citric acid is more than 50 mg/g;

the content of valine is more than 0.6mg/g, the content of tyrosine is more than 0.4mg/g, the content of proline is more than 0.2mg/g, and the content of phenylalanine is more than 0.2 mg/g;

the content of dipeptide in the small molecular polypeptide is more than 30mg/g, the content of tripeptide is more than 50mg/g, the content of tetrapeptide is more than 10mg/g, and the content of pentapeptide is more than 0.6 mg/g;

5. A probiotic post-biotic product according to any of claims 1 to 4, characterized in that the ratio of the number of colonies of Lactobacillus paracasei Probiosci-101 and Lactobacillus fermentum ProSci-602 is 2-3;

preferably, the ratio of the colony numbers of the lactobacillus paracasei Probiosci-101 and the lactobacillus fermentum procci-602 is 1.

6. A probiotic post-biotic product according to any of claims 1 to 5, characterised in that the culture medium used for the fermentation consists of, in mass percentages: 1.5 to 7 percent of full-fat bean powder, 11 to 18 percent of skim milk powder, 0.05 to 0.5 percent of sodium citrate and the balance of water.

7. Probiotic post-biotic preparation according to any of claims 1 to 6, characterized in that it is a cosmetic, preferably a functional cosmetic.

8. A method of preparing a probiotic post-biotic product according to any of claims 1 to 7, characterized in that it comprises: inoculating probiotic strains into the sterilized culture medium, fermenting at constant temperature until the pH is 4.5-4.6, inactivating, and drying to obtain probiotic post-production product.

9. Use of a probiotic post-biotic product according to any one of claims 1 to 7 and prepared according to the method of manufacture of claim 8 for promoting skin health.

10. Use according to claim 9, wherein said skin health promoting application includes, but is not limited to, any of the following:

1) Auxiliary prevention and/or improvement of facial skin condition;

the skin condition includes, but is not limited to, increasing facial elasticity, reducing facial wrinkles, increasing facial moisture content, reducing facial oiliness, or reducing facial melanin.